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长期冷冻保存不影响外周血造血干细胞移植物质量:对天然、短期和长期冷冻保存造血干细胞的比较研究。

Long-Term Cryopreservation Does Not Affect Quality of Peripheral Blood Stem Cell Grafts: A Comparative Study of Native, Short-Term and Long-Term Cryopreserved Haematopoietic Stem Cells.

机构信息

Department of Haematology and Oncology, University Hospital Pilsen, Pilsen, Czech Republic.

Biomedical Center, Faculty of Medicine in Pilsen, Charles University in Prague, Pilsen, Czech Republic.

出版信息

Cell Transplant. 2021 Jan-Dec;30:9636897211036004. doi: 10.1177/09636897211036004.

Abstract

Cryopreserved haematopoietic progenitor cells are used to restore autologous haematopoiesis after high dose chemotherapy. Although the cells are routinely stored for a long period, concerns remain about the maximum storage time and the possible negative effect of storage on their potency. We evaluated the effect of cryopreservation on the quality of peripheral stem cell grafts stored for a short (3 months) and a long (10 years) period and we compared it to native products.The viability of CD34+ cells remained unaffected during storage, the apoptotic cells were represented up to 10% and did not differ between groups. The clonogenic activity measured by ATP production has decreased with the length of storage (ATP/cell 1.28 nM in native vs. 0.63 in long term stored products, < 0.05). Only borderline changes without statistical significance were detected when examining mitochondrial and aldehyde dehydrogenase metabolic activity and intracellular pH, showing their good preservation during cell storage. Our experience demonstrates that cryostorage has no major negative effect on stem cell quality and potency, and therefore autologous stem cells can be stored safely for an extended period of at least 10 years. On the other hand, long term storage for 10 years and longer may lead to mild reduction of clonogenic capacity. When a sufficient dose of stem cells is infused, these changes will not have a clinical impact. However, in products stored beyond 10 years, especially when a low number of CD34+ cells is available, the quality of stem cell graft should be verified before infusion using the appropriate potency assays.

摘要

冷冻保存的造血祖细胞用于在高剂量化疗后恢复自体造血。尽管这些细胞通常被长期储存,但人们仍然对最长储存时间以及储存对其效力的可能负面影响感到担忧。我们评估了冷冻保存对短(3 个月)和长(10 年)期储存的外周造血干细胞移植物质量的影响,并将其与天然产物进行了比较。在储存过程中,CD34+细胞的活力保持不变,凋亡细胞占 10%,各组之间无差异。通过 ATP 产生测量的集落形成活性随着储存时间的延长而降低(天然产物中 ATP/细胞为 1.28 nM,长期储存产物中为 0.63,<0.05)。当检查线粒体和醛脱氢酶代谢活性以及细胞内 pH 时,仅检测到具有统计学意义的边缘变化,表明细胞储存过程中其保存良好。我们的经验表明,冷冻储存对干细胞质量和效力没有重大负面影响,因此可以安全地储存自体干细胞至少 10 年。另一方面,10 年以上的长期储存可能导致集落形成能力的轻微降低。当输注足够剂量的干细胞时,这些变化不会产生临床影响。然而,在储存超过 10 年的产品中,特别是当 CD34+细胞数量较少时,在输注之前应使用适当的效力测定法来验证干细胞移植物的质量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6fc4/8327011/6591c3ba4b90/10.1177_09636897211036004-fig1.jpg

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